Hi,
I am trying to develop an HPLC method for sucrose quantitation using UV detection (190-200 nm). Can someone please suggest a column for this application? I do not access to an RI detector; what can I do to enhance the sensitivity at 190-200 nm?

Thanks a lot for your help.

Suncrose and other sugars can be separated on an amino column using an acetonitrile / water mobile phase. I'd start at 70% MeCN and work my way up ti increase retention. Personally, I like the Asahipak NH2-P50 column available from a variety of suppliers. I have one from Keystone scientific that works well. Without derivitization, UV sensitivity of this component will be poor, if you see it at all. I would really suggest an RI or ELSD if at all possible

Hello,
you are going to make your life not easy without an RI detector. But, it is possible to detect at 190nm, if you don't have an RI detector. Mostly Amino NH2 columns are used. Reproducibility is not the best, but still the best choice if you have only a few samples. Polymer based NH2 columns are much better in terms of reproducibility, but price is much higher.
You have only a chance to get extrem small and symmetrical peaks with a spherical material 5µm!
Runtime on an 4,6x250 will be around 30 min, but with Acetonitrile/Water 75:25 you can separate Rhamnose, Xylose, Fructose, Glucose, Sucrose, Maltose, Trehalose, Lactose, Melibiose, Raffinose! Detection is UV at 190nm. I would highly recommend to use a thermostate (30 - 40°C)for your column. Flow rate should be 1ml/min.
If you like you can check some applications on our data base. Just click Tosohbiosep.com and go to Tosoh data base in Tokio. This might give you some ideas. But all applications are done with RI detection. It's many years ago I saw such an application with UV detection.
Good luck.
Gerhard Kratz

I will strongly recommend to use pure water as an eluent for low UV detection, as acetonitrile is difficult to obtain without absorbtion in low UV. Depending on other compounds in your sample, either reversed phase or ion exclusion columns can be used for separation of sucrose. To be able to suggest a specific column, I will need to know, what you want to separate the sucrose from.